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INTERNATIONAL RESEARCH JOURNAL OF PHARMACY www.irjponline.com ISSN 2230 – 8407

Research Article

ANTIOXIDANT EFFECTS OF LOVASTATIN ON COPPER-MEDIATED IN-VITRO OXIDATIVE MODIFICATION OF LDL IN INFLAMMATION INDUCED

HYPERLIPIDEMIA IN RATS Amir Khan1*, Shozab Jawed2, Mahesha Nand3, Lakhvinder Singh3, Rishab Jain and Munish Thakur5 1Department of Biotechnology & Biochemistry, Division of Life Science, Sardar Bhagwan Singh Post Graduate Institute

(SBSPGI) of Biomedical Sciences & Research Balawala, Dehradun- 248 161 UK, India 2Department of Biotechnology, Beehive College of Advance Studies, Dehradun, UK, India

3Department of bio-medical Sciences, Dolphin Post Graduate Institute of Bio-medical & Natural Sciences, Dehradun, UK, India

4Department of Biotechnology, Seth Jai Parkash Mukand Lal Institute of Engineering & Technology, (Chota Bans) Radaur, Yamuna Nagar (Haryana)

5Department of Pharmaceutical Chemistry, UCST, Dehradun, UK, India

Article Received on: 20/02/12 Revised on: 17/03/12 Approved for publication: 21/04/12 *Dr. Amir Khan, Assistant Professor, Department of Biochemistry, Sardar Bhagwan Singh Post Graduate Institute of Biomedical Sciences & Research Balawala, Dehradun, UK, India. E-mail: amiramu@gmail.com ABSTRACT Cardiovascular diseases (CVD) are the main cause of disability and premature dead worldwide. Epidemiological studies have suggested a link between atherosclerosis and inflammation. Atherosclerosis is a multifaceted diseases process with several different well defined risks factors, such as hypercholesterolemia, hypertension and diabetes. The present study was carried out to investigate the efficacy of antioxidant agent lovastatin. The study comprises the antioxidant status of Lovastatin by analyzing all the parameters in plasma, TL, TC, TG, VLDL-C, LDL-C, non-HDL-C, MDA and in-vitro oxidizability of LDL in absence or presence of Lovastatin. All the plasma lipids parameters, TL, TC, TG, VLDL-C, LDL-C, non-HDL-C and MDA levels were significantly increased in inflammation induced hyperlipidemic (IIH-C) rats. After 4-weeks administration of Lovastatin (1mg/ml) to IIH-C rats were useful in the prevention and treatment of inflammation induced hyperlipidemia, CVD and atherosclerosis. Lovastatin significantly reduced the overall oxidative burden and effectively ameliorated the above altered parameters. Keywords: Cardiovascular diseases (CVD), Atherosclerosis, Hypercholesterolemia, Hypertension and Inflammation. INTRODUCTION Cardiovascular disease (CVD) is the main cause of disability and premature death worldwide, and is projected to remain the leading cause of death. CVD is common in the general population, affecting the majority of adults. Hence, this disease greatly contributes to the rising costs of health care in the world. It is a major public-health challenge, especially in low and middle income countries. Excessive dietary lipids and cholesterol are the major factor of relevance for the development of hypertriglyceridemia and hypercholesterolemia, two important cardiovascular risk factors. Hyperlipidemia with accompanying increase in peripheral inflammation is a risk factor for stroke. Abnormalities in lipid profiles, folate metabolism and other traditional risk factors (e.g., diabetes mellitus and hypertension) play a rather peripheral role and serve to amplify the atherosclerotic process initiated by persistence of infection and inflammation1.Several clinical and epidemiological studies indicate that diabetes mellitus (DM) is an independent risk factor for CVD in both men and women, which increases the CVD risk by two-to six-fold relative to non diabetic subjects2,3. Infection and inflammation are accompanied by cytokine-induced alterations in lipid and lipoprotein metabolism. Of note, inflammatory cytokines are increased and play a pathogenic role in a variety of very common disorders, such as diabetes, obesity, metabolic syndrome, hypertension, chronic heart failure, chronic renal failure, and atherosclerosis4. Moreover, recent epidemiological studies have strongly suggested that

disorders that lead to systemic inflammation increases the risk of developing CVD. Studies have shown that patients with autoimmune diseases such as systemic lupus erythematosus, rheumatoid arthritis, or psoriasis have an increased risk of CAD5-8. Furthermore, a number of clinical disorders that are well recognized to increase the risk of CVD, such as diabetes, chronic renal failure and obesity, are now recognized to induce systemic inflammation9,10. Both infection and inflammation induce the systemic host response known as acute phase response (APR), and produce many abnormalities that could increase the risk of developing atherosclerosis including alterations in lipid and Lipoprotein metabolism, which is often, mediated by cytokines, particularly highly sensitive C-reactive protein, TNF-α, IL-1 and IL-6. Hyperlipidemia or hyperlipoproteinemia is the condition of abnormally elevated levels of any or all lipids and/or lipoproteins in the blood. It is the most common form of dyslipidemia (which also includes any decreased lipid levels). Lipids (fat-soluble molecules) are transported in a protein capsule, and the size of that capsule, or lipoprotein, determines its density. Lipid-lowering drugs such as statins have also been shown to antagonize inflammation. Lovastatin is a member of the drug class of statins, used for lowering cholesterol (hypolipidemic agent) in those with hypercholesterolemia and so preventing cardiovascular disease. Lovastatin is a naturally occurring drug found in food such as oyster mushrooms (Mevacor (Merck & Co.) in the United States Gunde-Cimerman N, Cimerman A. Mar 1995) and red yeast rice11. Lovastatin, a natural product with

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a powerful inhibitory effect on HMG-CoA reductase, were discovered in the 1970s, and taken into clinical development as potential drugs for lowering LDL cholesterol12,13. As discuss above, the role of inflammation in atherosclerosis has burgeoned. In this study we investigate the efficacy of antioxidant agent lovastatin by analyzing all the parameters in plasma, TC, VLDL-C, LDL-C, HDL-C and its sub fraction( HDL2-C, HDL3-C) ,Conjugated dine, MDA and as well as in-vitro oxidizability of Low Density Lipoprotein. MATERIAL AND METHODS Chemicals: 1-Chloro 2, 4-Dinitrobenzene was purchased from Central drug house, Pvt. Ltd. (India). All other chemicals used for this study were of analytical grade and obtained from HIMEDIA (India), Sisco (India), Ashirwad (India), Sigma-Aldrich (USA), Miles (USA), Acros (USA) and Lovastatin drug was supplied as a gift from Saimira Innoform Pvt. Ltd. Chennai, India. Estimation: Plasma triglyceride14, Plasma Cholesterol15,Plasma VLDL-C16, Fractionation of Plasma lipoprotein such as LDL17, HDL and its sub fractions-HDL2, HDL3

18, Plasma FRAP 19, ex vivo and in vitro Cu++-mediated LDL oxidation19,20. Experimental Design: The experimental (IAEC no-bc1962) study was approved by the Dolphin Institute of Biomedical and Natural Sciences, Dehradun, Uttarakhand, where the study was conducted. Healthy male albino rats, weighing about 150-180g were purchased from Indian Veterinary Research Institute, (IVRI) Bareilly (India), were maintained to animal house environmental condition prior to the experiment. For the present study, animals were divided into following 3 groups:-NC (normal control), IIH-C (inflammation induced hyperlipidemic control), and IIH-LT (inflammation induced hyperlipidemic Lovastatin treated). Diet/Drug Administration: The rats were given pelleted rat chow. Maintenance and treatment of all the animals was done in accordance with the principles of Institutional Animal Ethics Committee constituted as per the directions of the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), India. Six rats in IIH-LT group were given 1.0 mg Lovastatin/rat/day, through gastric intubation for 4 weeks. Induction of Inflammation: Inflammation was induced in IIH-C and IIH-LT by the subcutaneous injection of turpentine (0.5ml/rat) in the dorsolumbar region and left for five hours. Collection of Blood and Plasma: For the estimation of different parameters, overnight fasted rats in each group were anaesthetized and blood drawn from cardiac puncture, and were collected in heparinised tube. Plasma was separated from blood by centrifugation at 2500 rpm for 30 min. Statistical evaluation: This was done by employing two-tailed student t-test as describe by Bennet and Franklin (1967)21. P values less than 0.02 were considered significant. RESULTS Impacts of Lovastatin on average body weight in each group of rats: Table-1 depicts the average body weight (g) of N-C, IIH-C, IIH-LT was167g, 166g and 170g, whereas, the average body weight of N-C, IIH-C, IIH-LT rats showed a significant gain of 35%, 24% and 39% respectively after 4 weeks of treatment. These results demonstrate that in inflammation induced hyperlipidemic Lovastatin treated rats (IIH-LT) the gain in body weight after 4 weeks was significantly higher than N-C rats.

Table 1. Average body weight in each group of rats before and after 4 weeks of Lovastatin treatment Average body weight/rat (g)

Group Before treatment After Treatment

N-C 167.12±2.65*

226.75±14.62* (+35.68%)a

IIH-C 166.74±2.92* 208.13±16.13* (+24.82%)b

IIH-LT 170.21±6.967*

238.21±12.16* (+39.95%)a

*Values are mean ± SD from 6 rats in each group, N-C, normal control; IIH-C, inflammation induced hyperlipidemic rats; IIH-LT, fed 1 mg

Lovastatin/rat/day for 4 weeks., Significantly different from N-C at bp<0.001, Significantly different from IIH-C at ap<0.001.

Effects of Lovastatin on plasma total lipid (TL), triglycerides (TG) and total cholesterol (TC) in inflammation induced hyperlipidemic rats after 4 weeks of treatment: As seen in Fig 1, all the plasma lipids parameters were significantly increased in Inflammation induced hyperlipidemic (IIH-C) rats, when compared to N-C values. Total lipids (TL), triglycerides (TG) and total cholesterol (TC) significantly increased from 385, 48, and 74 mg/dl in N-C to 497, 98, and 141 mg/dl, respectively, in IIH-C group. After 4 weeks of Lovastatin treatment, levels of TL, TG, and TC were significantly decreased by 6.9 %, 42 %, and 37 %, respectively, when compared to corresponding N-C values. These results demonstrate that 4-week treatment of inflammation induced hyperlipidemic rats with 1.0 mg Lovastatin mediated a significant reduction in above lipid parameters.

Fig.1 Impact of Lovastatin on, plasma total lipid (TL), triglycerides

(TG) and total cholesterol (TC) in inflammation induced hyperlipidemic rats after 4 weeks of treatment. Significantly different from IIH-C at

bp<0.05. *Values are mean (mg/dl) ± SD from pooled plasma of 6 rats in each group. N-C, normal control; IIH-C, Inflammation induced hyperlipidemic rats; IIH-LT fed 1 mg Lovastatin/rat/day for 4 weeks. Significantly different from N-C

at ap < 0.001. Effects of Lovastatin on the plasma lipoprotein lipids and on the ratios of LDL-C/HDL-C and HDL-C/TC: As seen in Fig 2, plasma VLDL-C, LDL-C and non-HDL-cholesterol (non-HDL-C) levels were significantly increased from 8.9 mg/dl, 49 mg/dl and 59 mg/dl in N-C to 18 mg/dl (104%), 107 mg/dl (118 %) and 127 mg/dl (114 %) respectively in IIH-C. After 4 weeks of Lovastatin treatment, both VLDL-C, LDL-C and non-HDL-C levels showed a significant reduction 44 %, 50 % and 47 %, respectively, in IIH-LT. Whereas HDL-C, HDL2-C and HDL3-C levels were decreased from 17, 5 and 9 mg/dl in IIH-C to 15 mg/dl (11

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%), 3 mg/dl (28 %) and 9 mg/dl (4 %), respectively, in IIH-C values. After 4 weeks of Lovastatin treatment (IIH-LT) HDL-C, HDL2-C and HDL3-C levels showed a significant increase of 59 %, 124 % and 45 %, respectively, when compared to corresponding values in IIH-C. These results demonstrate that Lovastatin is effective in reducing VLDL-C and LDL-C levels. On the other hand, in comparison to IIH-C values, treatment of Inflammation induced hyperlipidemic rats with Lovastatin mediated a significantly higher increase in HDL-C, HDL2-C and HDL3-C concentration. On the other hand, LDL-C/HDL-C and HDL-C/TC ratios were calculated from the data presented in Table 2 and 3. LDL-C/HDL-C ratio was significantly increased from 2.87 in N-C to 7.13 (148 %) in IIH-C group, when compared to ratio in N-C. After 4 weeks of treatment, the increase in LDL-C/HDL-C ratio was significantly prevented and decreased to 2.00 in IIH-LT, which is close to normal control value. HDL-C/TC ratio was significantly decreased from 0.228 in N-C to 0.105 (53 %) in IIH-C group, as seen in Table 2. Lovastatin treatment to these rats significantly prevented the increase in HDL-C/TC ratios and fully restored them to a ratio value similar to N-C.

Fig 2 Impacts of Lovastatin on plasma lipoprotein lipids, in

inflammation induced hyperlipidemic rats after 4 weeks of treatment. *Values are mean (mg/dl) ± SD from pooled plasma of 6 rats in each group, N-C, normal control; IIH-C, Inflammation induced hyperlipidemic rats; IIH-LT fed 1 mg Lovastatin/rat/day for 4 weeks, Significantly different from N-C

at ap<0.001 and bp<0.05, Significantly different from IIH-C at ap<0.001.

Table 2. Impacts of Lovastatin on the ratio of LDL-C/HDL-C, HDL-C/TC, in inflammation induced hyperlipidemic rats after 4 weeks of

treatment

Parameter NC IIH-C IIH-LT

LDL-C/HDL-C

2.87±0.003*

7.13±0.012* (+148.43 %)a

2.20±0.005* (-69.14 %)a

HDL-C/TC 0.228±0.023*

0.105±0.002* (-53.39 %)a

0.269±0.016* (+156.19 %)a

*Values are mean (mg/dl) ± SD from pooled plasma of 6 rats in each group, N-C, normal control; IIH-C, Inflammation induced hyperlipidemic rats; IIH-LT fed 1 mg Lovastatin/rat/day for 4 weeks, Significantly different from N-C

at bp<0.001, Significantly different from IIH-C at ap<0.001. Impacts of Lovastatin on plasma total antioxidants and lipid peroxidation products: Fig 3 depicts the antioxidant impact of Lovastatin on plasma concentrations of total antioxidants, conjugated diene, lipid hydroperoxide and MDA in inflammation induced hyperlipidemic rats. In IIH-C rats, plasma total antioxidants level was reduced from a control value of 50 to 36 (27%) mmole/dl. Treatment of IIH-

LT rats with Lovastatin for 4 weeks resulted in a significant increase of total antioxidants levels by 15 % when compared to IIH-C value. The oxidative stress induced in IIH-C rats significantly enhanced plasma lipid peroxidation products, such as conjugated diene, lipid hydroperoxide and MDA. Formation of conjugated diene, lipid hydroperoxide and MDA in plasma was increased from 8.48, 1.12 and 1.18 in N-C to 12.35 (45 %), 1.89 (68 %) and 2.98 (152%) mmole/dl, respectively, in IIH-C. After Lovastatin treatment, in IIH-LT, a significant decrease of 15 %, 33 % and 33 % was seen in the formation of conjugated diene, lipid hydroperoxide and MDA, respectively, when compared to corresponding values in IIH-C rats. These results demonstrate that in IIH-C rats, due to increase in oxidative stress, total antioxidants level was decreased, whereas, concentration of plasma conjugated diene, lipid hydroperoxide and MDA were significantly increased. Lovastatin treatment significantly restored the total antioxidants level and blocked the increase in plasma conjugated diene, lipid hydroperoxide and MDA to a level close to corresponding normal values.

Fig.3 Antioxidant impact of Lovastatin on plasma total antioxidants,

conjugated diene (CD), lipid hydroperoxide (LHPO) and Malondialdehyde (MDA) contents in inflammation induced

hyperlipidemic rats after 4 weeks of treatment. *Values are mean (µmole/dl) ± SD from pooled plasma of 6 rats in each

group. N-C, normal control; IIH-C, Inflammation induced hyperlipidemic rats; IIH-LT fed 1 mg Lovastatin/rat/day for 4 weeks, significantly different

from N-C at ap<0.001., Significantly different from IIH-C at ap<0.001. Impacts of Lovastatin on the ex vivo and in vitro Cu++ mediated LDL Oxidation, conjugated diene formation, lag phase and total MDA release: Table-3 depicts the ex vivo base line diene conjugation (BDC) levels of LDL in inflammation induced hyperlipidemic rats (IIH-C) was increased by 54 % respectively, in comparison to the corresponding N-C values. Feeding of Lovastatin to inflammation induced hyperlipidemic rats partially blocked the in vivo oxidation of LDL and reduced their BDC levels by 20% respectively in comparison to the corresponding IIH-C values. As expected, the lag phase time of LDL oxidation was reduced from 90 min in N-C to 58 min in IIH-C. Treatment of IIH-LT rats with Lovastatin restored the lag phase time of LDL oxidation to 68 min. On the other hand the ex vivo base line levels of MDA in LDL was significantly increased by 55 % in (IIH-C) rats, when compared to corresponding values in normal control(N-C) rats. Lovastatin treatment significantly blocked the in vivo increase in the formation of MDA of LDL in inflammation induced

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hyperlipidemic rats and reduced their levels by 31%, in comparison to inflammation induced hyperlipidemic(IIH-C) rats. Similarly maximal in vitro oxidation of LDL was achieved after 12h of incubation with CuSo4 in each group.

The CD and MDA formation were significantly increased when compared to NC values, after 4 weeks of Lovastatin treatment, both values are significantly blocked.

Table 3 Ex-vivo and copper-mediated in-vitro oxidation of LDL, conjugated diene formation, lag phase and total MDA release in Inflammation

induced hyperlipidemic rats LDL-Oxidation

GROUP

Conjugate dine formation * MDA Content#

Basal Maximal Lag Phase$ Basal Maximal^

N-C 174.51 1038.32 90 +4.65±0.768

13.45±0.94

IIH-C 270.31+ (+54.89%)†

1425.47

(+37.29%)Ω 58

(- 35.55%) ¶

+7.25±0.134a

(+55.91%)‡

22.92±1.05a (+70.40%) Ω

IIH-LT 215.43+ (-20.30%) ††

1002.24

(29.70%) α 68

(+17.24%)§ +4.98±0.067a (-31.31%) ††

14.02±1.23a (-38.83%) α

+Values are mean ±SD from pooled plasma of 6 rats in each group. *conjugated diene values are expressed as nmole malondialdehyde equivalents/mg protein. Basal conjugated diene values represent the status of oxidized LDL in-vivo. $The lag phase defined as the interval between the intercept of the tangent of the slope of the curve with the time expressed in minutes. ^Maximal in vitro oxidation of LDL was achieved after 12h of incubation with CuSo4 in each group,

†Percent increase with respect to basal value in N-C, ††Percent decrease with respect to basal value in IIH-C, ¶ Percent decrease with respect to lag phase value in N-C,§ Percent increase with respect to lag phase value in IIH-C, Ω Percent increase with respect to maximal value in N-C, α Percent decrease with respect to

maximal value in IIH-C, Significantly different from N-C at aP<0.001. DISCUSSION The present study demonstrates the extensive proatherogenic changes, that occurred as a part of the host response to turpentine (acute localized sterile inflammation) administration, on a variety of parameters, like, plasma and lipoprotein lipids in plasma, liver lipid peroxidation products, ex vivo and in vitro oxidizability of LDL, erythrocytes MDA release; erythrocytes, liver and plasma total antioxidant. Pretreatment of stressed rats with Lovastatin significantly reduced the overall oxidative burden and effectively ameliorated the above altered parameters, thus, indicating a potent atheroprotective effect of Lovastatin. Our results demonstrate a significant increase in plasma total lipids and TG in turpentine (IIH-C) stressed rats. A similar increase in serum TG in LPS treated hamsters or rats were previously reported 22. In another report an increase in plasma TG level was seen during inflammation, induced by turpentine oil in pigs23. The increase in plasma TG levels is apparently due to an increase in VLDL which can be the result of either increased VLDL production or decreased VLDL clearance. These results are consistent with the finding that low doses of LPS stimulate VLDL production, whereas high doses of LPS inhibit VLDL clearance in rats22. A significant increase in plasma total cholesterol (TC) of IIH-C stressed rats was observed, which is in agreement with other reports, indicating an increase in serum TC level in LPS treated rodents, as well as in turpentine treated pigs. Similar to plasma lipids, VLDL-C, LDL-C and atherogenic non-HDL-C levels were also increased in stressed animals, indicating that the increase in plasma TC is apparently due to increase in VLDL-C and LDL-C concentrations. On the other hand, a low decrease of 12 % in plasma antiatherogenic HDL-C level of IIH-C rats, was seen, which is not comparable to a much higher decrease of 25 % and 45 % in HDL-C of LPS treated hamsters and turpentine treated pigs, respectively 24,23. Similar to plasma TG and TC in liver was also significantly increased in IIH-C rats. Therefore, tocotrienols may exert their cholesterol lowering effect in inflammation /infection induced hyperlipidemic rats in a similar manner as previously reported for hyperlipidemic animals [25,26] and humans27,28.

Mechanism wise, as previously shown in HepG2 cells, as well as in normolipidemic and hyperlipidemic rats, tocotrienols reduce cholesterol synthesis by suppressing HMG-CoA reductase activity, which in turn is reduced by a decline in its protein mass29. The decline in protein mass may be achieved by inhibition of HMG-CoA reductase synthesis and/or enhanced degradation. Consistent with in vivo results in rats25, γ-tocotrienol has been shown to mediate the suppression of enzymatic activity and protein mass of HMG-CoA reductase in HepG2 cells through decreased synthesis (57 % of control) and enhanced degradation (2.4-fold versus control) of the enzyme. In addition, γ-tocotrienol was shown to up regulate LDL receptor in mammalian cells and may be implicated in part for the reduction of apoB-lipoprotein in vivo 29. Thus, tocotrienols reduce cholesterol formation in mammalian cells by suppressing HMG-CoA reductase activity through two actions: decreasing the efficiency of translation of HMG-CoA reductase mRNA and increasing the controlled degradation of HMG-CoA reductase protein, post-transcriptionally. In addition, another report indicates that γ-tocotrienol influences apoB secretion by both cotranslational and posttranslational processes involving a decreased rate of apoB translocation and accelerated degradation of apoB in HepG2 cells. This activity correlated with a decrease in free and esterified cholesterol30. Taken together, the information indicates an association between the suppression of hepatic cholesterol synthesis and apoB secretion, and the observed lowering of apoB and LDL-C levels in animal and human models31. In our result, show that the ex vivo base line diene conjugation (BDC) levels of LDL in Inflammation induced hyperlipidemic (IIH-C) rats was increased by 54 % respectively, in comparison to the corresponding N-C values. Feeding of Lovastatin to Inflammation induced hyperlipidemic rats partially blocked the in vivo oxidation of LDL and reduced their BDC levels by 20 % respectively in comparison to the corresponding IIHC values. As expected, the lag phase time of LDL oxidation was reduced from 90 min in N-C to 58 min in IIHC. Treatment of IIH-LT rats with Lovastatin restored the lag phase time of LDL oxidation to 68 min. It has been established that LDL-C/HDL-C and HDL-

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C/TC ratios are good predictors for the presence and severity of CAD32-44. LDL-C/HDL-C and HDL-C/TC ratios were calculated from the data presented in Table 2 and 3. LDL-C/HDL-C ratio was significantly increased from 2.87 in N-C to 7.13 (148 %) in IIH-C group, when compared to ratio in N-C. After 4 weeks of treatment, the increase in LDL-C/HDL-C ratio was significantly prevented and decreased to 2.20 in IIH-LT, which is close to normal control value. On the other hand, HDL-C/TC ratio was significantly decreased from 0.228 in N-C to 0.105 (53 %) in IIH-C group. Lovastatin treatment to these rats significantly prevented the increase in HDL-C/TC ratios and fully restored them to a ratio value similar to N-C. In addition, the ratios related to HDL-C in Lovastatin treated rats were positively modulated and restored similar to normal control value, indicating normalization of cholesterol levels associated with the above lipoproteins Oral pretreatment of rats with Lovastatin for 28 days significantly prevented the turpentine induced adverse effects and ameliorated the levels of all the evaluated parameters. Our results strongly suggest that the alleviation of inflammatory conditions is due to potent lipid lowering and free radical scavenging properties of Lovastatin and, thus, can be useful in the therapy of systemic inflammatory process which might induce atherosclerosis. Based on these findings, the anti-inflammatory potential of Lovastatin looks promising and more comprehensive studies should be undertaken to determine their actual mode of action. In conclusion, considering the strong hyperlipidemic/athero-protective and antioxidant, and possibly anti-inflammatory actions of Lovastatin, intake of Lovastatin may be useful in the prevention and treatment of infection/inflammation induced hyperlipidemia and atherosclerosis. ACKNOWLEDGMENT The authors like to acknowledge University Grant Commission (UGC), New Delhi (India), for financial support. This study was carried out at the Department of Biochemistry, J N Medical College, Aligarh Muslim University, Aligarh, India. The authors like to thank Dr. Z. H. Beg, Er Shakir Khan, Nasir Khan for giving guidance from time to time and Dr. Asif Ali, Chairman, for providing facilities to carry out research work. REFERENCES 1. Mehta JL, Saldeen TGP, Rand K. Interactive Role of Infection,

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atherogenic sd-LDL in novice smokers. Journal of Pharmacy Research. 20114;(8): 2519-2522.

34. Chauhan S K, Thapliyal RP, Ojha S K, Rai Himanshu, Singh Puneet, Singh Manendra and Khan Amir. Antidiabetic and Antioxidant Effect of Ethanolic Root Extract of Boerhaavia diffusa in Streptatozocin-Induced Diabetic Rats. Journal of Pharmacy Research. 2011; 4(2): 446-448.

35. Chauhan S K, Thapliyal RP, Ojha S K, Rai Himanshu, Singh Puneet, Singh Manendra and Khan Amir. Hepatoprotective Role of Nigella sativa on Turpentine Oil Induced Acute Inflammation in Rats. International Journal of Chemical and Analytical Science. 20112; (8): 103-107.

36. Khan Amir, Chauhan S K, Singh R N and Thapliyal R P.

Chemotherapeutic Properties of Boerhaavia diffusa and Black Caraway Oil on DMBA-Induced Hypercholesterolemia in Animals. International Journal of Chemical and Analytical Science. 20112; (12): 1260-1264.

37. Khan Amir, Malhotra Deepti, Chandel Abhay S, Chhetri Samir, Fouzia, Rathore N S. In vitro Antioxidative effect of Boerhaavia diffusa on Copper mediated Oxidative Modification of LDL Ishaq in Type-II Diabetic patients. Pharma Science Monitor. 2011; p-1168-1181.

38. Khan Amir, Chandel Abhay Singh, Ishaq Fouzia, Chhetri Samir, Rawat Seema, Malhotra Deepti, Khan Salman and Rathore Nagendra Singh (2011). Protective Role of Dietary Tocotrienols and Black Caraway Oil on Infection and Inflammation Induced Lipoprotein Oxidation in Animals. Der Pharmacia Sinica. 2011; 2-2: 341-354.

39. Abhay Singh Chandel, Amir Khan, Fouzia Ishaq, Samir Chhetri, Seema Rawat and Deepti Malhotra (2011). Hypolipidemic effects and Antioxidant Activity of Tocotrienols on Cigarette Smoke Exposed Rats. Der Chemica Sinica. 2011; 2-2: 211-222.

40. Khan Amir, Chandel A S, Ishaq F, Rawat S, Chettri S, and Malhotra D (2011). Therapeutic Impacts of Tocotrienols and Boerhaavia diffusa on Cholesterol Dynamics, Lipid Hydroperoxidation and Antioxidant status on Hyperlipidemic Rats: Induced by Oxidized Cholesterol. Recent Research in Science & Technology. 2011; 3(11): 13-21.

41. 41.Chhetri Samir, Khan Amir, Rathore N S, Ishaq Fouzia, Chandel Abhay S, and Malhotra Deepti. Effect of Lovastatin on Lipoprotein Lipid Peroxidation and Antioxidant Status in Inflammation Induced Hyperlipidemic Rats. J of chemical and pharmaceutical research. 2011; 3(3): 52-63.

42. Khan M Salman, Khan Amir and Iqbal Jouhar, Effect of dietary tocotrienols on infection and inflammation induced lipoprotein oxidation in hamsters. 2011. International Journal of Pharmacy and PharmaceuticalSciences. 2011; 3(3): 277-284.

43. Khan Amir, and Malhotra Deepti. Protective Role of Ascorbic Acid (Vitamin C) Against Hyperlipidemia and Enhanced Oxidizability of Low Density Lipoprotein in Young Smokers. European Journal of Experimental Biology. 2011; 1 (1) :1-9

44. 44.Khan Amir, Ishaq Fouzia, Chauhan SK, Therapeutic Impacts of Tocotrienols (Tocomin) on Cholesterol Dynamics, and the ex vivo and Cu++ mediated in vitro oxidative modification of lipoprotein in rats, exposed to cigarette smoke. Drug Invention Today. 2012; 4(1): 314-320

45. Khan Amir, Ishaq F, Chandel Abhay S, and Chettri Samir (2012). Effect of dietary tocotrienols on antioxidant status and lipoprotein oxidation in rats, exposed to cigarette smoke. Asian Journal of Clinocal Nutrition. 2012; 1-12.

Source of support: Nil, Conflict of interest: None Declared